Expression of Inhibitory Molecules (Arginase-1, IDO, and PD-L1) by Myeloid-Derived Suppressor Cells in Multiple Myeloma Patients in Remission.

We studied suppressor potential of myeloid-derived suppressor cells (MDSC) in multiple myeloma patients, including before and after mobilization of hematopoietic stem cells (HSC), by evaluating the expression of arginase-1 (Arg1), indolamine-2,3-dioxygenase (IDO), and PD-L1 in MDSC subsets. The study included 20 multiple myeloma patients in remission, 5 patients with progression, as well as 10 sex-and age-matched healthy donors. The expression of Arg1, IDO, and PD-L1 in circulating granulocytic MDSC (G-MDSC, Lin-HLA-DR-CD33+CD66b+), monocytic MDSC (M-MDSC, CD14+HLA-DRlow/-), and early-stage MDSC (E-MDSC, Lin-HLA-DR-CD33+CD66b-) was evaluated by flow cytometry. Multiple myeloma patients in remission were characterized by reduced expression of Arg1 in M-MDSC in comparison with donors. The expression of Arg1 in M-MDSC depended on the number of induction therapy lines performed and was significantly lower in patients who received ⩾2 lines and responded with remission. Patients with multiple myeloma progression (resistant to therapy) showed significantly increased expression of Arg1 and PD-L1 in M-MDSC, as well as increased expression of Arg1 in E-MDSC. After G-CSF-induced mobilization of HSC, the content of circulating Arg1-expressing M-MDSC increased significantly. Considering the presence of MDSC in apheresis products, MDSC suppressive activity is discussed as a factor affecting the outcomes of autologous HSC transplantation in multiple myeloma patients.

The Cardio- and Neuroprotective Effects of Corvitin and 2-Oxoglutarate in Rats with Pituitrin-Isoproterenol-Induced Myocardial Damage.

Heart diseases, especially acute coronary syndrome (ACS), are among the most severe illnesses that often lead to death. Despite significant advances in the prevention and treatment of ACS, the incidence of the disease and its complications are very serious. The imbalance between pro- and antioxidant systems, the formation of active carbonyl compounds, and the end products of glycation in the blood and tissues are the key moments in the development of heart and neurological disorders leading to a change of behavioral responses. So, the search for antioxidants with cardio- and neuroprotective effects is an urgent task. This study was aimed at evaluating the effects of Corvitin and 2-oxoglutarate on physiological parameters, heart histology, and markers of carbonyl/oxidative stress of rats with pituitrin-isoproterenol-induced myocardial damage (PIMD). Increased sweating, tachycardia, significantly decreased locomotor and exploratory activity, changes of ECG, heart histology, and biochemical changes were observed in the PIMD-group. The administration of Corvitin or 2-OG led to the recovery of locomotor and cognitive activities of the rats, improvement in heart histology, a decrease in the levels of thiobarbituric acid reactive substances, advanced glycated end products, and various changes in the activity of the antioxidant enzymes, 6 days after PIMD. So, Corvitin and exogenous 2-OG show cardio- and neuroprotective effects through the decrease of carbonyl/oxidative stress and regulation of the activity of the antioxidant system.

DISTRIBUTION OF GLIAL FIBRILLARY ACIDIC PROTEIN IN DIFFERENT PARTS OF THE RAT BRAIN UNDER CADMIUM EXPOSURE.

The chronic effects of low doses of cadmium on the distribution of soluble and filament forms of glial fibrillary acidic protein (GFAP) and their polypeptide fragments in different parts of the rat brain were investigated. Obtained results showed dose-dependent effect of cadmium on the soluble form of GFAP and more pronounced effect on the filament form and composition of the polypeptide fragments of the protein in the rat brain. Prolonged intoxication by cadmium ions in a dose of 1.0 µg/kg of body weight induced a significant decrease in soluble GFAP and an increase in the filament form in the rat brain, pointing to the development of reactive astrogliosis and the risk of neurodegeneration.

Diet-induced changes in brain structure and behavior in old gerbils.

BACKGROUND/OBJECTIVES: Aging is associated with many physiological alterations such as changes in metabolism, food intake and brain dysfunction. Possible ways to correct age-related brain dysfunction using dietary treatments still remains undeveloped. The aim of our research was to investigate whether long-term dietary treatment with 2-oxoglutarate (2-OX), which is involved in many regulatory pathways, together with pancreatic-like enzymes of microbial origin (PLEM), which ensure appropriate digestion and absorption of nutrients, affects age-related changes in the brain morphology and cognitive function in old Mongolian gerbils. MATERIALS/METHODS: Experiment was comprised of two separate studies. Samples of the hippocampus were obtained from male Mongolian gerbils of different ages (n=63 in the first study, n=74 in the second study). Immunohistochemistry was used for visualization of the nestin/NeuN-positive neuronal progenitors. Changes in amount of neural cell adhesion molecules (NCAMs) were estimated using enzyme-linked immunosorbent assay. For assessment of cognitive and sensorimotor functions, the T-maze spontaneous alternation test and the adhesive removal test (ART) were used. The ultrastructure of the CA1 hippocampal area was visualized using transmission electron microscopy. RESULTS: Long-term treatment with 2-OX+PLEM led to a significantly increased amount of nestin/NeuN-positive cells in the CA1 hippocampal area and positive changes in learning and sensorimotor functions. As for synaptic transmission, changes in the spatial distribution of synaptic vesicles, as well as the redistribution of NCAM forms, were observed in the hippocampal synapses of the old gerbils. CONCLUSIONS: Taken together, our data show that dietary supplementation with 2-OX+PLEM not only enhances the proliferation and differentiation of neuronal progenitors, but also improves age-related deficits in the morphological and functional state of the brain of old gerbils. Thus, suggesting that a 2-OX+PLEM-enriched diet could also improve brain functions that have deteriorated with age.

Activity of trypsin-like enzymes and gelatinases in rats with doxorubicin cardiomyopathy.

Activity of trypsin-like enzymes (ATLE) and gelatinases A and B were studied in the blood plasma and extracts from cardiac muscle, cerebral cortex and cerebellum of rats with cardiomyopathy caused by anthracycline antibiotic doxorubicin against the background of preventive application of corvitin and α-ketoglutarate. ATLE significantly increased in blood plasma and extracts from cerebral cortex but decreased in extracts from cardiac muscle and cerebellum in doxorubicin cardiomyopathy (DCMP). In addition, a significant increase of activity of both gelatinases in plasma and tissue extracts was observed. Preventive administration of corvitin and α-ketoglutarate resulted in differently directed changes of activity of the above mentioned enzymes in heart and brain tissues. Obtained data confirm the hypothesis about activation of proteolysis under the influence of anthracycline antibiotics and testify to selective effect of corvitin and α-ketoglutarate on ATLE and gelatinases.

Exogenous pancreatic-like enzymes are recovered in the gut and improve growth of exocrine pancreatic insufficient pigs.

The exocrine pancreatic insufficient (EPI) pigs grow less due to different disturbances in feed digestion, absorption, and retention. Use of pancreatic-like enzymes of microbial origin in pigs may improve feed use and performance in slow-growing pigs. The aim was to study gut recovery and effectiveness of pancreatic-like enzymes of microbial origin supplementation on pig performance. Six male pigs 10 to 12 kg BW underwent pancreatic duct ligation surgery to induce total exocrine pancreatic insufficiency (EPI). Three cannulas to access the gastrointestinal tract content were installed in stomach, duodenum, and ileum in EPI pigs and in 3 control (healthy) pigs. One month after surgery, enzymes were given before feeding and digesta samples were collected for analyses. The BW of EPI pigs did not increase during 1 mo following surgery (11.7 vs. 11.6 kg BW); however, BW increased after 1 wk of enzyme supplementation (12.1 kg BW). Coefficient of fat and N absorption increased (P < 0.05) in EPI pigs after enzyme supplementation. Activity of amylase, lipase, and protease in chyme samples of EPI pigs was very low compared to controls. In EPI pigs after enzyme supplementation, amylase activity increased from 5.32 to 72.9 units/mL but remained lower than that of healthy pigs (162.7 units/mL). Lipase activity increased from 79.1 to 421.6 units/mL, which was similar to that of controls (507.3 units/mL). Proteolytic activity increased from 7.8 to 69.7 units/mL but still did not reach control pigs (164.3 units/mL). In conclusion, exogenous microbial enzymes mimic endogenous pancreatic enzymes being recovered along the lumen of the gastrointestinal tract. These enzymes might be a useful tool to stimulate growth of slower-growing pigs after the weaning period.

Behavioral changes in response to feeding pancreatic-like enzymes to exocrine pancreatic insufficiency pigs.

Behavioral changes during pancreatic enzyme therapy have never been studied. The present study investigated behavioral changes in exocrine pancreatic insufficiency (EPI) pigs when their feed was supplemented with pancreatic-like enzymes of microbial origin. A crossover design study was used to test the effect of enzyme supplementation in 2 × 4 EPI pigs that underwent pancreatic duct ligation (PDL). After 40 d of adaptation, the study commenced, comprising 2 control and 2 enzyme feeding periods of 10 d each in sequence. On days 7 and 10 of each experimental period, behavior was monitored for 24 h and feed consumption and BW were recorded. Behavioral observations focused on the pigs' activity-- lying down or passive, or sitting, or standing or active--and were expressed as percentage activity for 24 h. During the adaptation period, BW gain was completely inhibited after PDL whereas for the entire study period, the body weight increased from 10.5 ± 1.1 to 14.0 ± 1.4 kg (P < 0.01). Exocrine pancreatic insufficiency pigs were more active when fed the enzymes (21 vs. 18% per 24 h; P < 0.01). Microbial enzyme supplementation not only improved the growth of the EPI pigs but it also increased their activity. This behavior change contradicts the generally accepted norm that satiety evokes by digestion and subsequent nutrients absorption reduces human or animal motility.

The neuroprotective effect of 2-oxoglutarate in the experimental ischemia of hippocampus.

In this study we investigated the potential neuroprotective effect of 2-oxoglutarate (2-OG) on the hippocampus in the transient vessel occlusion ischemia model in the Mongolian gerbil. The morphological and biochemical studies were performed at 7 days after occlusion of carotid arteries. The acute reduction of NeuN-positive neurons in the CA1 pyramidal layer of the hippocampus was accompanied by increased staining intensity for GFAP-positive astrocytes, indicative of glial reaction. The neuron death in the CA1 area coincided with a strong 2.4 fold decrease in the membrane forms of neuronal cell adhesion molecules and elevated levels of astrocyte-specific proteins (soluble GFAP to 2,6 times; filament GFAP to 1,5 times; calcium-binding protein S-100b to 1,6 times). Treatment with 2-oxoglutarate (2.28 g/l drinking water) for between 7 and 21 days attenuated the neuronal death and reactive astrogliosis in this model of experimental ischemia by 20-50%. Our results suggest that 2-OG may prevent the disturbances of neural cells that usually take place during ischemic pathology.

[Ultrasound assessment of the effectiveness of chemoradiotherapy in cervical cancer].

The study involved 64 patients with verified cervical carcinoma, aged 23-79, who had received combined chemo-radiotherapy. Results were evaluated by ultrasound monitoring. Most modem methods were used via transabdominal and transrectal access. Tumor structure changed and size diminished following vascularization decrease step in step with combined chemoradiotherapy. Ultrasound in combination with 3D ultrasound - angiography provided objective data on tumor state at each stage of therapy.

[Thyroid diseases in reproductive-age women living in Shoria mountains].

AIM: To analyze prevalence and structure of thyroid pathology in women of reproductive age living in Shoria mountains. MATERIAL AND METHODS: The examination of 409 women aged 18 to 40 living in mountain Shoria included assessment of social status, heredity, actual nutrition, visit to endocrinologist, ultrasound investigation of the thyroid, tests for thyroid hormones (TTH, T3, T4) in the serum, antibodies to microsomal fractions, iodine concentrations in the morning urine. RESULTS: Significant prevalence of thyroid diseases (62.3%), high rate of diffuse nontoxic goiter of the second degree (18.0%), nodular nontoxic goiter (11.8%), autoimmune thyroiditis (6.3%), hypothyroidism (5.5%), low median of iodine excretion with urine in women refer mountain Shoria to the zone of goiter endemia. CONCLUSION: Among migrants thyroid morbidity is significantly higher than among the native population. Eradication of goiter endemia will promote reproductive health in this region.

[Dynamics of changes in the heparin-binding activity of rat brain proteins after fractionated exposure to ionizing radiation of low intensity]. / Dinamika izmenenii geparinsviazyvaiushchei aktivnosti belkov mozga krys posle fraktsionirovannogo vozdeistviia ioniziruiushchego izlucheniia nizkoi intensivnosti.

In experiment on white rats it was shown that fractionated exposure to 1 sGy/day during 25 days is followed by the directed change in the total heparin-binding activity of proteins in the rat brain (both in males and in females) with maximum deviation from control by the 7th day after accumulation of a total dose of 25 sGy. The diverse in hemisphere cortex and hippocampus were different from those in brain trunk regions and cerebellum. It is supposed that a thin overturning of the intercellular and "cell-matrix" interactions in CNS modulates compensation and adaptation processes under chronic X-ray irradiation with a small dose of low intensity due to regulation of the free/bound heparansulphate correlation.

[Change in heparin-binding activity of rat brain proteins after single exposure to ionizing radiation at a dose of 0.25 Gr]. / Izmenenie geparin-sviazyvaiushchei aktivnosti belkov mozga krys posle odnokratnogo vozdeistviia ioniziruiushchego izlucheniia v doze 0.25 Gr.

Sex-dependent description of the total protein heparin-binding activity in different rat brain regions was established in experiment on white rats. In norm named parameter above in males, than in females on a 22%. The specific alteration of the total heparine-binding activity was found in cortex, cerebellum, middle brain, hippocampus, striatum and pons 1, 12, 24, 120 and 168 hours after whole body single exposure to ionizing radiation with a dose of 0.25 Gy. The most pronounced changes were typical for limbic system. In the cortex and cerebellum of males no significant changes of the activity were found. The data suggest that modification of the system of heparin/heparin-binding proteins, which takes part in regulation of the intercellular brain communication, occurs at early hours after exposure to ionizing radiation.

[Efferent methods in the therapy of puerperal and postoperative complications]. / Efferentnye metody terapii poslerodovykh i posleoperatsionnykh oslozhnenii.

Ninety-six patients with chorioamnionitis in labor were treated. Clinical observations and laboratory findings demonstrated high efficiency of multiple-modality treatment including use of efferent methods: plasmapheresis, incubation of cell mass with antibiotics, and ultra-violet exposure of the blood.

The influence of low doses 131I-induced maternal hypothyroidism on the development of rat embryos.

The aim of our research was to create and verify a model for studying the effects of a low dose of 131I and 131I-induced maternal hypothyroidism on the development of the embryo's thyroid gland and brain. The given dose (150 microCi) corresponds to the absorbed dose of 0.5 Gy. This dose is similar to that dose received by large numbers of the population of the C.I.S. regions polluted by radioactive isotopes of iodine as a result of the Chernobyl accident in 1985. Thirty-five female Wistar rats and their 168 newborn pups were used for observation. The females were divided into a control group and four experimental groups (each distinguished by the time of 131I injection: group I - no less than 12 days before mating; groups II, III and IV - on 5th, 10th and 16th days of gestation, respectively). In all the experimental female groups the incorporate dose of 131I led to hypothyroidism accompanied by a 43% reduction in the thyroxin level and by a nearly 8-fold increase in the TSH level. However, the influence of maternal hypothyroidism on the development of the thyroid gland and brain of embryos depends on the time when 131I took effect. There is a reduction in the weight of the newborns' brain and thyroid gland, total body mass. The hormonal status of the newborns' thyroid gland also changes. The proposed model will allow us to study many aspects of induced changes in the brain and thyroid gland of the embryos which develop under conditions of maternal hypothyroidism resulting from a low dose of 131I, administered at the critical times of development.

[Isolation and purification of a membrane hyaluronate-binding protein from embryonic human brain]. / Vydelenie i ochistka membrannogo gialuronatsviazyvaiushcego belka iz mozga émbrionov cheloveka.

A membrane hyaluronate-binding protein from cerebral cortex of human embryonic brain (22-24 weeks) was purified by affinity, ion exchange chromatographies and gel-filtration. While gel-filtration analysis the protein had Mm 250 kDa. Electrophoresis under reduction conditions in the presence of DS-Na revealed a major band with Mm 85 kDa and two minor binds with Mm 68 and 36 kDa. The isolated protein did not react with antibodies against known hyaluronate-binding and other proteins with similar mass. The results show that a new membrane hyaluronate-binding protein was isolated and purified from human embryonic brain.

Some morphological changes in the rat thyroid gland during experimental hyperphenylalaninemia.

BACKGROUND: Little is known about the effect of phenylketonuria on the thyroid gland. In the present study, this problem was investigated by using a defined experimental model of hyperphenylalaninemia (HPA). METHODS: The experimental group was subjected to an HPA regimen (Matsuo and Hommes, 1988. Neurochem. Res., 13:867-870) from the 5th day of postnatal development. The pups were decapitated on the 7th, 14th, 21st, 28th, and 35th days. The thyroid glands were fixed in Bouin's fluid and routinely embedded in paraffin. The staining techniques used were Mallory-Slinchenko's method, toluidin blue, silver impregnation of the basement membrane, immunohistochemical staining of the proliferating cell nuclear antigen (PCNA), and neuron-specific enolase (NSE). RESULTS: The size of the follicles was less than that in the control group. There were no substantial changes in the epitheliomer structures. In almost all of the treated groups, a reduction in the number of PCNA+, NSE+, and mast cells was observed until the 28th day. On the 28th day of HPA, the level of mast cell degranulation was higher (61%) than that in the control group. On the 35th day, these parameters began to reach normal levels. From the 28th day, degenerative changes in the thyroid glands of treated animals were observed in the NSE+ cells. CONCLUSIONS: The HPA condition mainly has an influence on the number and structure of the NSE+ cells of the thyroid gland. One may assume that under HPA the increase in mast cell degranulation plays a significant role in the normalisation of the parameter of the thyroid gland.

Effect of experimental hyperphenylalaninemia on the postnatal rat brain.

The molecular mechanism of the disturbance of brain development caused by phenylketonuria remains mostly unknown. We have studied three molecular markers that reflect the development of neurons, glia and the extracellular matrix of the postnatal rat brain in an animal model of hyperphenylalaninemia, in order to elucidate the possible mechanism by which increased phenylalanine influences brain development. The content of NCAM, GFAP and hyaluronate-binding activity were compared in cerebellum and telencephalon of normal rats and those subjected to high phenylalanine. No statistically significant changes were found in telencephalon when experimental animals were compared to controls. In the hyperphenylalaninemic cerebellum, the developmental dynamic of NCAM content (represented by two peaks at about postnatal days 5 and 22 during normal development) is dramatically altered. The GFAP content in the cerebellum of treated rats exceeded those in controls significantly during late developmental stages (postnatal days 28-35). Hyaluronate-binding activity in the extracellular protein fraction from treated rat cerebellum was increased compared to normal rat at the early stages of development only (postnatal day 7). These results suggest that high serum phenylalanine may lead to permanent brain dysfunction through a disturbance of a wide range of developmental events.

Neural cell adhesion molecule (N-CAM) distribution may predict the effect of neurotoxins on the brain.

The neural cell adhesion molecule (N-CAM) is a convenient neurospecific marker for investigating the effects of neurotoxins on cell migration, cell recognition and differentiation of neurons during development. In this report, we discuss the developmental toxicity of valproic acid studied by two different approaches (the immunochemical detection of N-CAM content and polypeptide composition, and immunohistochemical analysis of N-CAM topography). Immunohistochemical analysis of distribution of N-CAM as a surface marker on the neural cells predicted the effect of the neurotoxin.